The <i>cop</i> operon is required for copper homeostasis and contributes to virulence in <i>Streptococcus pneumoniae</i>

Shafeeq, Sulman; Yeşilkaya, Hasan; Kloosterman, Tomas G.; Narayanan, Geetha; Wandel, Michal P.; Andrew, Peter W.; Kuipers, Oscar P.; Morrissey, Julie A.

doi:10.1111/j.1365-2958.2011.07758.x

Cited by 149 publications

(229 citation statements)

References 57 publications

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“…4C). It was shown recently that this operon could be transcriptionally induced by, and is sensitive to, toxic levels of copper sulfate (Shafeeq et al 2011), a phenotype which we confirmed (data not shown). Since S. pneumoniae is unlikely to encounter such nonphysiological levels of copper, our findings suggest that, instead, the operon is used to counter imbalances in divalent cation concentrations.…”

Section: Validation Of In Vitro Genotype-phenotype Relationships Leadsupporting

confidence: 78%

A fine scale phenotype–genotype virulence map of a bacterial pathogen

2012

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A large fraction of the genes from sequenced organisms are of unknown function. This limits biological insight, and for pathogenic microorganisms hampers the development of new approaches to battle infections. There is thus a great need for novel strategies that link genotypes to phenotypes for microorganisms. We describe a high-throughput strategy based on the method Tn-seq that can be applied to any genetically manipulatable microorganism. By screening 17 in vitro and two in vivo (carriage and infection) conditions for the pathogen Streptococcus pneumoniae, we create a resource consisting of >1800 interactions that is rich in new genotype-phenotype relationships. We describe genes that are involved in differential carbon source utilization in the host, as well as genes that are involved both in virulence and in resistance against specific in vitro stresses, thereby revealing selection pressures that the pathogen experiences in vivo. We reveal the secondary response to an antibiotic, including a dual role efflux pump also involved in resistance to pH stress. Through genetic-interaction mapping and gene-expression analysis we define the mechanism of attenuation and the regulatory relationship between a two-component system and a core biosynthetic pathway specific to microorganisms. Thus, we have generated a resource that provides detailed insight into the biology and virulence of S. pneumoniae and provided a road map for similar discovery in other microorganisms.

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Section: Validation Of In Vitro Genotype-phenotype Relationships Leadsupporting

confidence: 78%

A fine scale phenotype–genotype virulence map of a bacterial pathogen

2012

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“…It is established that copper is an effective antimicrobial agent ex vivo, and there is evidence that it may play a similar role in vivo. Pathogens that lack copper efflux systems colonize poorly in the mammalian lung, possibly because of an inability to detoxify increased levels of copper in the host tissue (21). Detoxification of intracellular copper is achieved by efflux via P-type ATPase and RND (resistancenodulation-cell division) family efflux systems (22).…”

Section: Discussionmentioning

confidence: 99%

Global discovery of colonization determinants in the squid symbiontVibrio fischeri

Brooks

Gyllborg

Cronin

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

105

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Animal epithelial tissue becomes reproducibly colonized by specific environmental bacteria. The bacteria (microbiota) perform critical functions for the host's tissue development, immune system development, and nutrition; yet the processes by which bacterial diversity in the environment is selected to assemble the correct communities in the host are unclear. To understand the molecular determinants of microbiota selection, we examined colonization of a simplified model in which the light organ of Euprymna scolopes squid is colonized exclusively by Vibrio fischeri bacteria. We applied highthroughput insertion sequencing to identify which bacterial genes are required during host colonization. A library of over 41,000 unique transposon insertions was analyzed before and after colonization of 1,500 squid hatchlings. Mutants that were reproducibly depleted following squid colonization represented 380 genes, including 37 that encode known colonization factors. Validation of select mutants in defined competitions against the wild-type strain identified nine mutants that exhibited a reproducible colonization defect. Some of the colonization factors identified included genes predicted to influence copper regulation and secretion. Other mutants exhibited defects in biofilm development, which is required for aggregation in host mucus and initiation of colonization. Biofilm formation in culture and in vivo was abolished in a strain lacking the cytoplasmic chaperone DnaJ, suggesting an important role for protein quality control during the elaboration of bacterial biofilm in the context of an intact host immune system. Overall these data suggest that cellular stress responses and biofilm regulation are critical processes underlying the reproducible colonization of animal hosts by specific microbial symbionts.H umans and other animals are often sterile before birth, from which point they immediately proceed to acquire environmental bacteria (1). The bacteria that reproducibly colonize animal hosts are critical for host tissue development, immune system development, and nutrient acquisition. The selection process by which the functional symbionts take residence in the animal from among the great diversity of environmental microbes is poorly understood, so model systems have been especially valuable to examine how specific patterns of colonization are shaped by the genetic makeup of the bacteria and the host environment (2).The light organ of the Hawaiian bobtail squid, Euprymna scolopes, is colonized exclusively by the Gram-negative luminous bacterium Vibrio fischeri. The host inhabits seawater containing 10 6 bacteria per milliliter, with V. fischeri comprising at most 0.02% of the environmental population (3). E. scolopes hatch without symbionts, but then rapidly acquire environmental bacteria and proceed to select for V. fischeri in a "winnowing" process that ensures colonization by only the specific symbiont (4). The squid-Vibrio system thus presents an opportunity to investigate the processes that underlie acquisition of s...

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“…The pathogenic bacterium Streptococcus pneumoniae contains the operon copY-cupA-copA, which encodes the repressor CopY, a protein of unknown function (CupA), and the copper-ATPase CopA (Shafeeq et al, 2011). Transcription of the cop operon is induced by copper and repressed by zinc, suggesting that the two metals compete for the same ligands in CopY.…”

Section: Streptococcus Mutans and Streptococcus Pneumoniaementioning

confidence: 99%

“…CopA is the major copper tolerance determinant in Streptococcus pneumoniae, while CupA is less important for copper defence. In addition to its role in copper homeostasis under free-living conditions, the cop operon is important for virulence (Shafeeq et al, 2011). Expression of the cop operon …”

Section: Streptococcus Mutans and Streptococcus Pneumoniaementioning

confidence: 99%